Revealing Seasonal Dietary Niche Overlap Among Sympatric Large Carnivores using DNA Metabarcoding

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Patterson, Stéphanie Périquet-Pearce, Madeline H. Melton, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7990153/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 15 Mar, 2026 Read the published version in Scientific Reports → Version 1 posted 11 You are reading this latest preprint version Abstract Understanding how large carnivores partition dietary resources is essential for assessing intra-guild competition and informing conservation strategies. In this study, we used DNA metabarcoding of scats to quantify and compare the diets of sympatric African lions ( Panthera leo ) and spotted hyenas ( Crocuta crocuta ) across wet and dry seasons in the Greater Etosha Landscape of Namibia. Across 98 scat samples, we identified 19 vertebrate prey species. Overall, large ungulates dominated both carnivores’ diets. For lions, the most frequent prey items included gemsbok ( Oryx gazella ), common eland ( Taurotragus oryx ), plains zebra ( Equus quagga burchelli ), and blue wildebeest ( Connochaetus taurinus ). For spotted hyenas, the most frequent prey items were plains zebra, gemsbok, springbok ( Antidorcas marsupialis ), and black rhinoceros ( Diceros bicornis bicornis ). Despite differing dietary niche breadth, with lions exhibiting the broadest across both seasons, diet composition was similar between species and seasons. These results confirm a high diet overlap and limited resource partitioning both within and among these large carnivore species across seasons, likely facilitated by opportunistic scavenging and kleptoparasitism. Both species exhibited broader dietary niche breadths during the wet season, likely reflecting increased prey availability and dispersion. Ongoing monitoring of carnivore diets using molecular tools, which provides a more accurate and comprehensive identification of diet items than manual sorting, will be essential for detecting changes in resource use and interspecific interactions in response to shifting environmental conditions and anthropogenic pressures. Biological sciences/Ecology Earth and environmental sciences/Ecology Biological sciences/Zoology African lions diet composition dietary niche breadth interspecific competition resource partitioning spotted hyenas Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Large carnivores play critical roles in maintaining ecosystem health and diversity (del Rio et al., 2001 ). Yet, despite multifaceted conservation initiatives, large carnivore populations are declining globally due to anthropogenic pressures (Parmesan, 2006 ; Di Marco et al., 2014 ; Ripple et al., 2014 ) including human-mediated conflicts due to their large home range requirements and the expanding human footprint (Carbone and Gittleman, 2002 ). As the human populations footprint expands, large carnivores are becoming increasingly restricted to protected areas (Mills and Harvey, 1998; Riggio et al., 2012), which can lead to higher levels of interspecific interactions and intra-guild competition. Thus, monitoring large carnivore populations, especially where intra-guild competition is likely to be high, is imperative for conservation efforts (Linnell and Strand, 2000 ; Burgar et al., 2019 ). In regions with well-preserved predator guilds, large carnivore species often occupy overlapping geographic ranges and compete for prey resources. Interspecific competition can drive community structure and function, and affect the distribution, demographic patterns, and behavioral strategies of co-occurring species (Chase et al. 2002 , Chesson and Kuang 2008 , Vanak et al., 2013 ). Because large carnivore populations are primarily shaped by the abundance and availability of their main food items (Karanth et al., 2004 ; Hayward & Kerley, 2008 ), prey represents a critical resource that coexisting carnivores can partition to reduce competition (Schoener, 1983 ; Hayward & Kerley, 2008 ). Intra-guild competition intensity can vary spatio-temporally due to changes in prey availability, which can fluctuate due to a multitude of biotic and abiotic factors such as seasonality, droughts, climate change, or disease outbreaks (Walther et al., 2002 ; Duncan et al., 2012 ; Zidon et al., 2017 ). While numerous studies have investigated carnivore competition and resource partitioning, data is lacking in certain areas of sub-Saharan Africa, where largely intact carnivore guilds exist. Understanding how sympatric carnivores partition dietary resources in these systems is thus essential to understand carnivore ecology (Radloff and du Toit, 2004 ) and inform conservation practices through vegetation and prey species management. Some of the most diverse and multifaceted terrestrial assemblages of large carnivores occur in sub-Saharan Africa, with several species of large carnivores occurring sympatrically including African lions ( Panthera leo ) (hereafter lions), spotted hyenas ( Crocuta Crocuta ), cheetahs ( Acinonyx jubatus ), leopards ( Panthera pardus ), and brown hyenas ( Parahyaena brunnea ). Lions and spotted hyenas are two of the most numerous large carnivores in this region where they overlap extensively in distribution (Hatton et al., 2015 ) and diel activity (Mills & Biggs, 1993 ; Hayward & Hayward, 2007 ; Patterson et al., 2024 ). To facilitate coexistence, lions and spotted hyenas have evolved disparate behavioral, social, and foraging strategies (Périquet et al., 2015 ; Swanson et al., 2016 ). Lions are ambush predators, who hide under vegetation and pursue medium-large ungulate prey (Hayward and Kerley, 2005 ; Périquet et al., 2015 ), yet also hunt in open areas (Stander, 1992 ) and scavenge opportunistically (Moleón et al., 2015 ; Amorós et al., 2020 ). Conversely, spotted hyenas are cursorial predators, typically chasing prey and using endurance for hunting success (Kruuk, 1972 ; Mills, 1990 ) but also readily scavenge carcasses when available (Watts & Holekamp, 2009 ; Moleón et al., 2015 ). Additionally, spotted hyenas do successfully hunt calves or juvenile large ungulate species (Kruuk, 1972 ; Holekamp et al., 1997 ; Salnicki et al., 2001 ). Lions and spotted hyenas engage in kleptoparasitism; although spotted hyenas lose more of their kills to lions than lions lose to spotted hyenas (Périquet et al., 2015 ), though this kind of intra-guild competition is highly variable based on pride and clan size and structure (Trinkel and Kastberger, 2005 ). With overlapping diel activity patterns, kleptoparasitism, and opportunistic scavenging, dietary overlap is expected to be high between lions and spotted hyenas with substantial variation across their shared geographic range. Previous studies report broad dietary niches for spotted hyenas (Vissia et al., 2023 ), though they have been found to have similar (Hayward, 2006 ) or narrower (Hayward & Kerley, 2008 ) niche breadths compared to lions. However, spotted hyenas may reduce their niche overlap by preying on different prey size via different sex or age classes (Périquet et al., 2015 ). Additionally, top prey items vary across studies for both species (e.g., Hayward, 2006 ), and the role of seasonality in shaping dietary niche breadth and overlap remains poorly understood (Vissia et al., 2023 ). Few studies have employed high-throughput sequencing methods like DNA metabarcoding to address these questions. DNA metabarcoding has been shown to have higher sensitivity and greater taxonomic resolution relative to conventional dietary studies that rely on morphological identification of undigested remains in scats (Stein et al., 2014 ; Galan et al., 2017). Morphological identification can introduce additional biases into dietary results through misidentification of closely related prey items, lack of detection for prey items with highly digestible tissues, and underestimating prey items (Monterosso et al., 2019). The conflicting findings in previous dietary studies, along with the limited application of molecular techniques, underscore the importance of assessing carnivore diet partitioning, especially between lions and spotted hyenas, to quantify dietary overlap in a rich carnivore community. Moreover, while little is known about the effects of seasonality on dietary niche overlap among large carnivores, understanding these dynamics is critical for advancing our knowledge of intra-guild resource partitioning and coexistence in shared landscapes. Our goal in this study was to use DNA metabarcoding to quantify dietary patterns of sympatric lions and spotted hyenas in a dryland ecosystem in southern Africa. Specifically, our objectives were to 1) quantify the vertebrate diet composition of sympatric lions and spotted hyenas; 2) quantify seasonal variation in diet composition within and between each species; 3) assess dietary niche breadth for each species across seasons; and 4) evaluate interspecific dietary overlap to determine if there is evidence of potential niche partitioning and competition. We also formulated a series of predictions. First, we predicted that both species would have higher diversity in their diet during the wet season due to increased availability of young herbivores during the birthing season for many ungulate species, and animals dispersing across the landscape and away from predictable waterhole locations, forcing carnivores to be less selective for prey species. Second, based on scavenging activity and previous studies (Hayward, 2006 ; Vissia et al., 2023 ), we predicted that spotted hyenas would have higher dietary niche breadths than lions during both seasons. Lastly, we predicted that both species would have high dietary overlap during both seasons because of kleptoparasitism and opportunistic scavenging. Methods Study Area This study was conducted in the Greater Etosha Landscape (GEL) encompassing Etosha National Park (“Etosha”), the Ongava Game Reserve (“Ongava”), and King Nehale communal conservancy (“King Nehale”) in north-central Namibia (Fig. 1 ) (Turner et al., 2022 ). Etosha is a 22,900 km 2 fenced protected area, although the fence is permeable to wildlife (Naha et al., 2023 ; Melton, 2024 ). Ongava is a fenced 300 km 2 private game reserve on the southern border of Etosha. King Nehale is a 508 km 2 communal conservancy located in the Oshikoto region, bordering the northeast corner of Etosha. One main source of income for people living in the conservancy is keeping livestock, including cattle, goats, and sheep (Nakanyala et al., 2022 ). The GEL is located within the semi-arid weather zone with an average annual rainfall of 380 mm (De Beer et al., 2006 ; Atlas of Namibia, 2022). Despite fencing around Etosha, the prey and predator guild composition in all these areas is very similar and together they form part of the same larger scale ecosystem, including movement of carnivores between Ongava and Etosha, and more so between King Nehale and Etosha (Melton, 2024 ). The main vegetation types in Etosha are grasslands, steppe, shrubland, Mopane ( Colophospermum mopane ) tree savanna, and mixed tree savanna (Le Roux et al., 1988 ). The landcover on Ongava is classified as Karstveld, with Mopane shrubland and woodland being the dominant vegetation type (~ 70%) and other areas with savannah-like vegetation (~ 30%, Berry and Loutit, 2002 ). During the wet season (November-April), rainfall provides additional temporal water sources, aside from the permanent artificial water sources across the landscape (Engert, 1997 ; Berezin et al., 2023 ). Both Etosha and Ongava are tourist destinations for wildlife-based viewing and support a broad diversity of carnivore and ungulate species [e.g., black-faced impala ( Aepyceros melampus petersi ), springbok ( Antidorcas marsupialis ), blue wildebeest ( Connochaetus taurinus ), plains zebra ( Equus quagga burchelli ), gemsbok ( Oryx gazella )], including megaherbivores [e.g, African elephant ( Loxodonta africana ), southern Angolan giraffe ( Giraffa giraffa angolensis ), southern black rhinoceros ( Diceros bicornis bicornis )]. However, prey species’ composition and abundance differ between Ongava and Etosha, and across the different regions of Etosha (west, central, and east) (Trinkel, 2010 ; Kilian and Kolberg, 2015 ). Sample Collection We collected 164 dried or fresh fecal samples [spotted hyena (n = 32) and lion (n = 132)] from October 2021 through March 2024 around Etosha, Ongava, and King Nehale. When divided by location, we collected 72 samples in Etosha [spotted hyena (n = 28) and lion (n = 44)], three samples in King Nehale [spotted hyena (n = 2) and lion (n = 1)], and 88 samples in Ongava [spotted hyena (n = 3) and lion (n = 82)]. Most samples (n = 140) were collected opportunistically from the ground while driving along roads, around waterholes, and near carnivore kill sites within our study area (approved by Namibian Ministry of Environment, Forestry, and Tourism under permit #AN202101004). For these samples, whole fecal samples were collected into a paper bag and dried naturally before being subsampled for DNA extraction, which included both the external surface and internal matrix to capture both consumer and prey DNA. Additionally, 24 samples [lions (n = 19), spotted hyenas (n = 5)] were collected during carnivore immobilizations as part of ongoing research (Appendix S1; Text S1). These samples were manually retrieved from the immobilized animal, placed into an open plastic vial and stored in a paper bag for natural drying. Due to the small amount collected directly from immobilized animals, these samples were not subsampled. All immobilizations were performed by veterinarians registered with the Namibian Veterinary Council and the Ministry of Environment, Forestry, and Tourism and were approved by the Namibian Ministry of Environment, Forestry, and Tourism (permit #AN202101004) as well as the University of Georgia Institutional Animal Care And Use Committee under protocols A2024 05-009-06 and A2021 04-013-Y3-A11. Laboratory Analyses and Data Curation All samples were shipped from Namibia to the United States in July 2024 and DNA was extracted from feces using DNeasy PowerSoil Pro DNA Kits (QIAGEN) in August 2025. Jonah Ventures, LLC conducted DNA amplification, sequencing, and bioinformatics for DNA metabarcoding (Appendix 1; Text S2). This technique analyzes DNA within scats using high-throughput sequencing using small, highly variable universal primers to identify prey species and confirm predator species (Pompanon et al., 2012 ; Xiong et al., 2017). To characterize prey DNA from carnivore fecal samples, we amplified a portion of the 12S rRNA gene using the primer pair 12SVertF (forward 5′- ACTGGGATTAGATACCCYACTATG − 3′) and 12SVertR (reverse 5′- GAGRRTGACGGGCGGTDT − 3′) (Evans et al., 2016 ). Both forward and reverse primers contained a 5’ adaptor sequence to allow for subsequent indexing and Illumina sequencing. Exact sequence variants (ESVs) were identified using the UNOISE3 algorithm (Edgar, 2016a ) implemented in VSEARCH (Rogens et al., 2016), retaining only those with ≥ 4 read counts and ≤ 1 expected error. We assigned taxa of ESVs using a custom reference database consisting of publicly available sequences on NCBI GenBank (Benson et al., 2005 ) and Jonah Ventures voucher sequences records, accepting the reference taxonomy for any taxonomic level with > 90% agreement within 1% of the top hits. We discarded any reads of taxa not occurring in the study region, and to limit the effects of contamination, only prey species that consisted of more than 1% of total prey sequences were retained (Shively et al., 2025 ). Because host DNA typically dominates the sequencing dataset (Forin-Wiart et al., 2018 ), we were able to confirm predator species for each sample. Data Analyses We conducted all analyses and produced figures using R version 4.1.1 (R Core Team, 2024 ). We summarized the data with both frequency of occurrence (FOO) and relative read abundance (RRA) at the species level, and calculated prey species richness for each carnivore species by season (wet and dry). To calculate FOO, we divided the number of occurrences of a prey species by the total number of carnivore scat samples and expressed FOO as a percentage. We calculated RRA for each prey species as the read count (the number of DNA sequence reads) for that the given prey species divided by the total number of reads in the carnivore scat sample, also expressed as a percentage. To examine diet composition (beta-diversity) of vertebrate prey species, we conducted separate three-dimensional non-metric multidimensional scaling (NMDS) analyses with 1000 permutations to visualize trends in variation between seasons and carnivore species. We used two complementary approaches to evaluate dietary similarity between and within carnivore species. First, we applied a non-parametric Analysis of Similarities (ANOSIM) with binary Jaccard dissimilarities to test whether prey composition within carnivore scats differed between lions and hyenas within each season. Because ANOSIM evaluates overall compositional differences but does not quantify the degree of overlap, we also calculated Pianka’s index of niche overlap (Pianka, 1973 ) based on FOO. To account for unequal sample sizes, we generated 95% bootstrap confidence intervals by stratified resampling of carnivore scats for both species (2,000 iterations). Together, these analyses allowed us to assess both whether diets differed significantly between carnivore species and the extent of dietary niche overlap, providing a more complete picture of dietary similarity across seasons. We used the vegan package in R (Oksanen et al., 2025 ) to conduct the NMDS and ANOSIM and spaa package in R (Zhang, 2016 ) to determine Pianka’s index. We calculated dietary niche breadth ( Ba ) for each carnivore species in both seasons by using Hurlbert’s standardized version of Levin's measure (Hurlbert, 1978 ; Smith, 1982 ) for FOO proportions of prey. To account for differences in sample size across carnivore species and seasons, we applied a bootstrap approach with 2,000 iterations. For each iteration, all samples were resampled with replacement within each carnivore species–season group, and standardized niche breadth was recalculated. Broader dietary niche breadth is indicated by a higher Ba while a narrow dietary niche breadth is indicated by a low Ba (ranging from 0–1). We did this overall with all data pooled, and separately for each study site [Etosha (including King Nehale) versus Ongava]. To evaluate completeness of our sampling efforts and prey richness, we generated sample-based rarefaction and extrapolation curves for each carnivore species and season using the specaccum function in the vegan package (Oksanen et al., 2022). Scat samples were converted to presence–absence matrices of prey taxa, and 1,000 random permutations were used to estimate mean prey richness and associated 95% confidence intervals. We further estimated asymptotic richness with the Chao1 estimator (specpool function) and plotted these estimates (± standard error) alongside rarefaction curves to assess whether observed prey detections approached estimated richness. For all analyses we pooled data from all study sites to generate broad inferences about carnivore diet, and we also visually examined diet composition between Etosha and King Nehale versus Ongava. Results Diet overview From the 164 samples that were sequenced, 98 had detectable vertebrate prey items and were included in analysis [spotted hyena (n = 29; dry season = 17, wet season = 12) and lion (n = 69; dry season = 54, wet season = 15)]. Overall, we detected 9 families and 17 genera encompassing 19 vertebrate prey species (Table 1 ). For spotted hyenas, the total read count was 127640 with a mean read count of 3868 per sample (± 4461 SD). For lions, the total read count was 107965 with a mean read count of 1367 per sample (± 2733 SD). Table 1 Percent frequency of occurrence (%FOO; number of fecal samples containing each prey species divided by total number of fecal samples and expressed as a percentage) and Relative Read Abundance (RRA%; total number of reads of each vertebrate prey species divided by total number of vertebrate reads and expressed as a percentage) of vertebrate species (12S rRNA) detected in the diet of African lions ( Panthera leo ) and spotted hyenas ( Crocuta crocuta ) based on season (dry = May-October; wet = November-April) in the Greater Etosha Landscape; “n” represents number of samples containing that prey species for that carnivore species Carnivore - Season Prey Species n Average RRA (%) FOO (%) Lion – Dry Baboon 1 7.63 1.69 Black rhinoceros 2 7.63 3.39 Black-faced impala 1 4.99 1.69 Blue wildebeest 8 7.32 13.56 Common eland 8 6.86 13.56 Gemsbok 15 7.37 25.42 Angolan giraffe 3 7.08 5.08 Hare spp. 2 4.29 3.39 Greater kudu 3 7.63 5.08 Mountain zebra 2 7.63 3.39 Plains zebra 6 6.42 10.17 Red hartebeest 2 3.95 3.39 Springbok 4 5.93 6.78 Turner's thick-toed gecko 1 7.63 1.69 White rhinoceros 1 7.63 1.69 Lion – Wet Black-faced impala 2 14.46 10 Blue wildebeest 2 3.94 10 Common eland 3 12.01 15 Gemsbok 3 13.90 15 Angolan giraffe 2 14.46 10 Hare spp. 2 7.32 10 Greater kudu 1 9.75 5 Plains zebra 3 9.76 15 Python spp. 1 4.71 5 Red hartebeest 1 9.69 5 Spotted Hyena - Dry Black rhinoceros 1 12.5 5.88 Black-faced impala 2 12.5 11.76 Cow 1 12.5 5.88 Common eland 1 12.5 5.88 Gemsbok 3 12.5 17.65 Greater kudu 1 12.5 5.88 Plains zebra 5 12.5 29.41 Springbok 3 12.5 17.65 Spotted Hyena - Wet Black rhinoceros 2 3.09 12.5 Cow 1 0.24 6.25 Common eland 1 15.82 6.25 Gemsbok 3 12.1 18.75 Angolan giraffe 2 13.07 12.5 Goat 2 15.7 12.5 Greater kudu 1 10.04 6.25 Plains zebra 3 14.13 18.75 Springbok 1 15.82 6.25 Spotted hyena diet From 29 sequenced spotted hyena samples, plains zebra was the most frequently consumed prey species in the dry season (FOO = 29.4%) followed by gemsbok ( Oryx gazella ) and springbok (FOO = 17.7% each). In the wet season, spotted hyenas equally consumed plains zebras and gemsbok (FOO = 18.8% each, Fig. 2 ). Overall, prey species richness was 10 and included two domestic livestock species, cow ( Bos taurus ; n = 2) and goat ( Capra hircus ; n = 2). Both domestic prey species were found in samples near or outside of northeast Etosha (Fig. 1 ). ANOSIM revealed no seasonal differences in diet composition for spotted hyenas (Table 2 ). Table 2 ANOSIM results comparing prey composition between African lions ( Panthera leo ) and spotted hyenas ( Crocuta crocuta ) in the Greater Etosha Landscape. Comparisons include between seasons (wet = November – April; dry = May – October) within each species, seasons between species, and overall between species across all samples. R-statistic and p-values are provided for each comparison Seasonality within species (wet vs.dry) R-stat p-value Spotted Hyena -0.024 0.73 Lion -0.008 0.58 Seasonality between species Dry 0.021 0.17 Wet -0.018 0.67 Overall between species Lion vs. Spotted Hyena 0.014 0.17 When we separated out by study site (Etosha and King Nehale versus Ongava), during the dry season, plains zebra and springbok were the most consumed prey items in Etosha (Fig. S1 ) while gemsbok and kudu ( Tragelaphus strepsiceros ) were the only consumed prey items in Ongava (Fig. S2 ). During the wet season, spotted hyenas primarily consumed plains zebra and gemsbok in Etosha (Fig. S1 ). Lion diet From 69 sequenced samples, lions consumed a wide variety of species but primarily large ungulates such as gemsbok (FOO = 25.4%), followed by blue wildebeest and eland ( Taurotragus oryx ) (FOO = 13.6% each), then plains zebra (FOO = 10.2%) during the dry season. In the wet season lions consumed eland, gemsbok, and plains zebra equally (FOO = 15% each), followed by black-faced impala, blue wildebeest, giraffe, and hare species (FOO = 10% each, Fig. 2 ). Overall, prey richness was 16 and ANOSIM revealed no significant seasonal differences in diet composition for lions (Table 2 ). Separated out by study site, during the dry season, lions consumed springbok most frequently in Etosha, followed by giraffe, plains zebra, eland, red hartebeest, and black rhinoceros equally (Fig. S1 ). During the dry season on Ongava, lions consumed gemsbok most frequently, followed by blue wildebeest, eland, and plains zebra (Fig. S2 ). During the wet season in Etosha, lions primarily consumed giraffe and plains zebra equally followed by eland, red hartebeest, blue wildebeest, kudu, and python species equally (Fig. S1 ). During the wet season on Ongava, lions, consumed gemsbok, eland, black-faced impala, and hare spp. equally, followed by blue wildebeest and plains zebra (Fig. S2 ). Dietary niche breadth Overall, during the wet and dry seasons, lions exhibited broader dietary niche breadth than spotted hyenas [lions ( Ba : dry = 0.40, wet = 0.45); hyenas ( Ba : dry = 0.28, wet = 0.38)]. Both species had broader dietary niche breadths during the wet season compared to the dry season (Fig. 3 ). In Etosha, lions exhibited broader dietary niche breadth than spotted hyenas in the dry season ( Ba : lions = 0.5, hyenas = 0.29), but spotted hyenas had slightly broader dietary niche breadths in the wet season ( Ba : lions = 0.47, hyenas = 0.44). Lions had broader dietary niche breadths in the dry season, while spotted hyenas were broader in the wet season (Fig. S3). In Ongava, lions had similar dietary niche breadths across seasons ( Ba : dry = 0.417, wet = 0.414), and broader dietary niche breadth compared to spotted hyenas in the dry season ( Ba = 0.09) (Fig. S4). Interspecific dietary variation We found no differences in dietary composition for either carnivore species overall (all data pooled) or across season. The NMDS ordination for dietary overlap (Fig. 4 ) had a stress value of 0 in the wet season and < 0.001 in the dry season. While a stress value of or near 0 is uncommon, it can occur in small or low-dimensional datasets but is under the threshold value (stress 0.05) in diet composition between lions and spotted hyenas within either season or overall. R statistics were close to zero or negative, indicating strong dietary overlap among these carnivores across seasons and overall (Table 2 ). Pianka’s index revealed moderate dietary overlap between lions and spotted hyenas overall (O = 0.75). Seasonal analyses showed that overlap between species was slightly higher in the dry season (O = 0.68) than in the wet season (O = 0.66). Within species, lions exhibited greater dietary similarity across wet and dry seasons (O = 0.84) compared to spotted hyenas (O = 0.77) (Table 3 ). Table 3 Pianka’s index of dietary overlap (O) between African lions ( Panthera leo ) and spotted hyenas ( Crocuta crocuta ). Overlap values are shown with 95% confidence intervals (CIs) based on bootstrap resampling (2,000 iterations). Results are presented for seasonal comparisons within species (wet vs. dry), between species within each season, and overall between species across all samples Seasonality within species (wet vs.dry) O 95% CI Spotted Hyena 0.77 0.34–0.85 Lion 0.84 0.53–0.88 Seasonality between species Dry 0.68 0.36–0.80 Wet 0.66 0.31–0.74 Overall between species Lion vs. Spotted Hyena 0.75 0.54–0.83 Rarefaction curves Rarefaction curves revealed that prey species richness in lion scats approached asymptotic levels, particularly in the dry season, where observed richness (15 taxa) closely matched Chao1 estimates (17.0 ± 2.6). In the wet season, lion richness was lower (10 taxa, 11.0 ± 1.7), suggesting some rare prey may not have been captured. Spotted hyena curves indicated lower observed richness overall (dry = 8, wet = 9 taxa), with estimates suggesting additional undetected prey species, particularly in the dry season (Chao1 = 15.5 ± 11.0), though uncertainty was high due to small sample size (Fig. 5 ). Discussion Determining how large carnivores partition dietary resources is critical for understanding intra-guild interactions and coexistence. Using DNA metabarcoding, we characterized the diets of lions and spotted hyenas in the GEL and found high dietary overlap between both species, regardless of the season. Despite differences in prey species richness and niche breadth between carnivore diets, there were no significant seasonal or interspecific differences in prey species composition. These findings suggest that in this landscape, resource partitioning among large carnivores may occur by targeting different sex or age classes of prey items or through mechanisms other than diet. Collectively, these data contribute to our overall understanding of dietary patterns of sympatric lions and spotted hyenas in a semi-arid ecosystem. Species with broad dietary niche breadths are typically considered dietary generalists while those with narrow dietary niche breadths are considered specialists (Carvalho & Cardoso, 2020 ). In our study, lions had a wider dietary niche breadth during both seasons compared to spotted hyenas; however, our findings demonstrate that lions and spotted hyenas exhibit broadly similar diets with consistently high overlap, suggesting that these two apex predators rely heavily on a shared prey base. When comparing dietary niche breadth between species, lions and spotted hyenas are reported to have similar dietary niche breadths, which aligns with our results (Hayward, 2006 ; Hayward and Kerley, 2008 , reviewed in Périquet et al., 2015 ). The comparison between seasons supported our prediction that both lions and spotted hyenas had broader dietary niche breadths in the wet season compared to the dry. This difference in seasonality has been previously documented in African mammal diets (Vissia et al., 2023 ) perhaps because some prey species are more abundant and more easily accessible (Trinkel, 2013 ; Pereira et al., 2014 ) during the birthing season or because rainfall leads to prey dispersing across the landscape. As a result, predators can no longer rely on high prey densities near predictable waterhole locations and may be less selective, consuming whatever prey they encounter. The results confirm our prediction that there would be high dietary overlap between both carnivore species, likely enhanced by kleptoparasitism and scavenging behavior. The overall Pianka’s index value (0.75) indicates substantial dietary similarity, and this pattern was maintained across seasons, with only minor fluctuations between the wet and dry periods. Apart from domestic livestock species, all prey species detected in the spotted hyena diet were also found in the lion diet. Extensive dietary overlap may indicate occurrences of kleptoparasitism by lions and spotted hyenas, which previously have been reported (Trinkel and Kastberger, 2005 ). Additionally, lions and spotted hyenas are documented competitors that consume similar prey items (Hayward, 2006 ; Hayward and Kerley, 2008 ) and exhibit comparable temporal activity patterns (Mills & Biggs, 1993 ; Hayward & Hayward, 2007 ; Patterson et al., 2024 ). Thus, our findings indicate that lions and spotted hyenas in Etosha may partition their prey resources by age or sex class, or spatially to co-exist (Patterson et al., 2025 ) and we recommend further studies to quantify prey partitioning across demographic classes and spatial scales to better understand mechanisms of niche differentiation and coexistence among large carnivores. In a review of spotted hyena diet across several countries, Hayward ( 2006 ) determined that gemsbok was the most frequently consumed prey species where it occurred, and plains zebra and springbok were typically consumed infrequently. Conversely, other studies (Gasaway et al., 1991 ; Trinkel, 2010 ) found plains zebra and springbok were the main prey species for spotted hyenas in some areas of Etosha, while Berry ( 1981 ) determined wildebeest were the main prey species in Etosha. Fester et al. ( 2021 ) found gemsbok and springbok were the main prey items in the Namib desert of southwest Namibia. In our study, spotted hyenas most frequently consumed plains zebras, gemsbok, and springbok, respectively. As such, spotted hyenas may target gemsbok but also consume plains zebras and springbok in the GEL due to their high abundance and availability compared to other ungulates. For example, in 2015, the Etosha density of gemsbok was estimated at 0.22 individuals/km 2 while plains zebra was 0.77 individuals/km 2 and springbok were 0.56 individuals/km 2 (Kilian and Kolberg, 2015 ). As plains zebra and springbok are grazers (Kingdon, 2015 ), and gemsbok are mixed feeders that primarily graze (Sponheimer et al., 2003 ; Lehmann et al., 2020 ), these results also align with Patterson et al. ( 2025 ) who found that spotted hyenas in Etosha select for areas of high grass cover during both seasons. Collectively, these studies indicate that in the GEL, spotted hyenas are likely hunting in open, grassy areas and primarily consuming grazing prey species. The range of prey items for lions showed no strong selection for either browsers or grazers, as both (browsers: giraffe; mixed: eland, gemsbok; grazers: plains zebra, blue wildebeest, Kingdon, 2015 ) were found as lion top prey species. Our results align with findings from Hayward and Kerley ( 2005 ) who found lions’ primary prey species were gemsbok, blue wildebeest, giraffe, eland, and zebra across several countries and Berry ( 1981 ) who found lions’ primary prey items in Etosha were gemsbok, wildebeest, and plains zebra. Consistent with other findings, lions opt for large prey species but will also consume small prey opportunistically (Hayward and Kerley, 2005 ; Davidson et al., 2013 ; Barnardo et al., 2020 ). This supports the optimal foraging theory, which suggests that a predator can distinguish among prey of differing profitability and typically select the most profitable types (Krebs, 1978 ). Large-bodied grazers were consumed more during the dry season in our study, which may indicate that lions were opting for higher caloric returns from large prey species to sustain themselves during higher periods of stress (Hayward and Kerley, 2005 ). Additionally, in the dry season herbivores congregate at fixed waterhole locations in the GEL, increasing predator access to a higher density of potential prey. Conversely, in the wet season lions appear to consume fewer species, with several at higher frequencies relative to the dry season (e.g., hare spp., black-faced impala, giraffe), potentially reflecting opportunistic foraging or scavenging due to greater prey dispersion. We found three spotted hyena samples with domestic livestock species (cow and goat). All samples were collected near or outside of Etosha in the northeast area (King Nehale communal conservancy) where livestock are abundant and spotted hyenas often leave the park and enter onto communal land (Naha et al., 2023 ; Melton, 2024 ; Patterson et al., 2025 ), sometimes resulting in retaliatory killings of carnivores after livestock depredation events (Stander, 2004 ; Goelst, 2018 , Naha et al. 2025 ). Unfortunately, there is little information on the extent and demography of spotted hyenas killed in anthropogenic areas surrounding Etosha, but local farmers report frequent sightings of spotted hyenas on their farms (Lendelvo et al., 2019 ). These findings contribute to our overall understanding of livestock depredation in a high human-carnivore conflict area. The rarefaction analyses highlight important differences in sampling completeness between lions and spotted hyenas, which help contextualize niche overlap between lions and spotted hyenas, as well as niche breadth estimates. We could not detect prey items in 66 samples due to predator DNA swamping, which occurs when host DNA is present in much higher concentrations than prey DNA (Shi et al., 2021 ). For lions, both wet- and dry-season rarefaction curves approached asymptotic levels, and Chao1 estimates suggested that only a small number of prey species were likely undetected, indicating that our dietary characterization for lions is robust and provides a reliable picture of seasonal prey use. In contrast, the rarefaction curve for spotted hyenas showed lower observed richness and substantially higher Chao1 estimates, particularly in the dry season where uncertainty in prey species detection was large, reflecting potential under-sampling. This limitation suggests that while lions and spotted hyenas appear to have high dietary overlap, the true extent of spotted hyena prey use may be broader than captured in this study. Consequently, the high dietary overlap we observed may partially reflect more complete sampling of lion diets compared to spotted hyenas. Nonetheless, the combination of dietary niche overlap and breadth results indicates that both species rely heavily on shared prey resources, with lions exhibiting more stable and well-characterized prey use across seasons. Future work that includes expanded spotted hyena sampling could help clarify whether spotted hyena dietary breadth converges more strongly with or diverges from that of lions under different seasonal conditions. We found notable differences in frequently consumed prey species and dietary niche breadth for both lions and spotted hyenas between study sites, likely reflecting variation in prey abundance and community composition within the sampled areas. Although both Etosha and Ongava are protected areas, they are managed by different entities and are different sizes, which may influence wildlife distributions and prey availability. However, the small number of spotted hyena samples from Ongava (n = 2) limited our ability to accurately characterize their dietary niche breadth at that site. Additionally, because sampling did not encompass the entire extent of Etosha, we were unable to provide a comprehensive synthesis of lion and spotted hyena diets across the entire GEL. Despite these limitations, this study provides valuable insights into the diets of lions and spotted hyenas where dietary data, particularly derived from high-resolution DNA metabarcoding, are lacking. Our study contributes to the area of emerging research using DNA metabarcoding analysis to compare diets of large, sympatric carnivores. The results revealed strong dietary overlap and limited niche partitioning among lions and spotted hyenas across both wet and dry seasons. Our findings suggest that prey availability and opportunistic feeding behavior may be the primary drivers of diet composition in this well-preserved carnivore guild. Despite differences in dietary niche breadth, both species showed high overlap in key prey taxa, perhaps due to opportunistic scavenging and kleptoparasitism. These findings emphasize the importance of maintaining abundant ungulate populations to support diverse predator communities. Importantly, the detection of domestic livestock in spotted hyena diets highlights the potential for human–wildlife conflict, emphasizing the need for continued management interventions around Etosha. Further diet monitoring is essential for detecting shifts in predator–prey dynamics in response to environmental change, prey population fluctuations, or increased anthropogenic pressures. Future research should also explore the spatial and temporal aspects of scavenging and interspecific interactions to better inform adaptive management and conservation strategies in protected areas and surrounding multi-use landscapes. Declarations Competing interests: The authors have no relevant financial or non-financial interests to disclose. Funding: This work was supported by University of Georgia and the US Department of Energy Office of Environmental Management Award Number DE-EM0005228 to the University of Georgia Research Foundation. Disclaimer: This manuscript was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information disclosed, or represents that its use not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not constitute or imply its endorsement, recommendation, or favoring by the United States Government or any agency thereof. The views and opinions of the authors expressed herein do not necessarily state or reflect those of the United States Government or any agency thereof. Author Contribution Jessica Patterson, James Beasley, and Stephanie Periquet-Pearce contributed to the study conception and design. Material preparation and data collection were performed by all authors. Date analyses and the first draft of the manuscript were completed by Jessica Patterson. All authors commented on previous versions of the manuscript and approved the final manuscript. Acknowledgement We thank the Ongava Game Reserve for providing some equipment and staff assistance. We thank the field technicians for their work in helping to collect and process scat/fecal samples. Thank you to the University of Georgia and the US Department of Energy Office of Environmental Management for funding assistance. Data Availability The datasets generated and analysed during the current study are available in the NCBI Sequence Read Archive repository at [https://www.ncbi.nlm.nih.gov/sra/PRJNA1357869](https:/www.ncbi.nlm.nih.gov/sra/PRJNA1357869) . References Amorós, M., Gil‐Sánchez, J.M., López‐Pastor, B.D.L.N. and Moleón, M. 2020. Hyaenas and lions: how the largest African carnivores interact at carcasses. Oikos , 129(12): 1820-1832. Atlas of Namibia Team. 2022. 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Peirson","email":"","orcid":"","institution":"Northern Michigan University","correspondingAuthor":false,"prefix":"","firstName":"Laura","middleName":"E.","lastName":"Peirson","suffix":""},{"id":550023763,"identity":"00ef16db-8dfa-4974-9e12-857937068c53","order_by":8,"name":"Diana J.R. Lafferty","email":"","orcid":"","institution":"Northern Michigan University","correspondingAuthor":false,"prefix":"","firstName":"Diana","middleName":"J.R.","lastName":"Laf","suffix":"J.R."},{"id":550023764,"identity":"c72fd987-c821-420a-8c85-783c3939fe4f","order_by":9,"name":"James C. 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16:04:46","extension":"html","order_by":16,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":185469,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/eb2549ec188c62f366fce24a.html"},{"id":96917934,"identity":"720358a5-1cc0-447c-ae62-7a29fcaeb862","added_by":"auto","created_at":"2025-11-27 14:10:46","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":771278,"visible":true,"origin":"","legend":"\u003cp\u003eLocation of collected fecal samples from African lions (\u003cem\u003ePanthera leo\u003c/em\u003e, red dots) and spotted hyenas (\u003cem\u003eCrocuta Crocuta\u003c/em\u003e, black dots) in the Greater Etosha Landscape, Namibia. Collection occurred between October 2021 and March 2024\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/c8f43627bfb0b42577c770d2.png"},{"id":96918374,"identity":"cd281f88-35e9-4414-a15e-24a6fa213404","added_by":"auto","created_at":"2025-11-27 14:11:51","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":150166,"visible":true,"origin":"","legend":"\u003cp\u003eSeasonal composition of vertebrate prey species detected in the diets of African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) in the Greater Etosha Landscape based on DNA metabarcoding. Bars represent the proportion of fecal samples in which each prey species was detected, calculated as the number of unique samples per species and season divided by the total number of samples for that carnivore and season. Results are shown separately for the dry season (green) and wet season (grey), and prey species are ordered by overall frequency of occurrence from left to right. Only prey species with ≥1% relative read abundance were included\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/c82a533242723fd358437ec4.png"},{"id":96842392,"identity":"5e42fac0-b5af-43be-8f67-4e9113f46830","added_by":"auto","created_at":"2025-11-26 16:04:45","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":45438,"visible":true,"origin":"","legend":"\u003cp\u003eSeasonal variation in standardized niche breadth (Hurlbert’s B′) for African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) in the Greater Etosha Landscape, in both the dry (May-October) and wet (November-April) seasons. Bars represent mean dietary breadth based on frequency of occurrence (FOO) of prey species in fecal samples, with error bars showing 95% confidence intervals from 2,000 bootstrap iterations\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/f13f51a913a86553df9c7d55.png"},{"id":96842393,"identity":"82aba46b-55cd-45f9-bf61-8a771c05a478","added_by":"auto","created_at":"2025-11-26 16:04:45","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":85422,"visible":true,"origin":"","legend":"\u003cp\u003eNon-metric multidimensional scaling plot for vertebrate species (12S rRNA) detected in the diet of African lions (\u003cem\u003ePanthera leo\u003c/em\u003e), and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) in the Greater Etosha Landscape, by season (dry = May-October and wet = November-April)\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/9ba551b62a0b46081c2a89a8.png"},{"id":96842396,"identity":"427becf6-0ee2-481a-8fbd-00e62ec98d84","added_by":"auto","created_at":"2025-11-26 16:04:45","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":55355,"visible":true,"origin":"","legend":"\u003cp\u003eSample-based rarefaction and extrapolation curves of prey species richness detected in lion (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyena (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) scats during dry (green) and wet (black) seasons. Solid lines represent mean richness estimates with shaded areas indicating 95% confidence intervals based on 1,000 random permutations. Triangles show observed richness, and vertical error bars represent Chao1 asymptotic richness estimates (± SE)\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/5afd21d00360fee4fb1727f2.png"},{"id":104739643,"identity":"45b6c418-6f8d-435d-92d3-f566d98c5d80","added_by":"auto","created_at":"2026-03-16 16:11:31","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2126777,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/691d8ee5-7b2a-47c8-ad45-5164ecc3b052.pdf"},{"id":96842398,"identity":"e4d60a8a-0d48-4e4d-ad60-11ff757caa9d","added_by":"auto","created_at":"2025-11-26 16:04:45","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":411910,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryFigures.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/9f1fd059b412c116d92657c9.pdf"},{"id":96918469,"identity":"4b6d1935-cb6c-4c1a-8cd1-51132e3680e8","added_by":"auto","created_at":"2025-11-27 14:11:58","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":107947,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryText.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7990153/v1/136497a1c0bdffe9d09d521b.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Revealing Seasonal Dietary Niche Overlap Among Sympatric Large Carnivores using DNA Metabarcoding","fulltext":[{"header":"Introduction","content":"\u003cp\u003eLarge carnivores play critical roles in maintaining ecosystem health and diversity (del Rio et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Yet, despite multifaceted conservation initiatives, large carnivore populations are declining globally due to anthropogenic pressures (Parmesan, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Di Marco et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Ripple et al., \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) including human-mediated conflicts due to their large home range requirements and the expanding human footprint (Carbone and Gittleman, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). As the human populations footprint expands, large carnivores are becoming increasingly restricted to protected areas (Mills and Harvey, 1998; Riggio et al., 2012), which can lead to higher levels of interspecific interactions and intra-guild competition. Thus, monitoring large carnivore populations, especially where intra-guild competition is likely to be high, is imperative for conservation efforts (Linnell and Strand, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Burgar et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn regions with well-preserved predator guilds, large carnivore species often occupy overlapping geographic ranges and compete for prey resources. Interspecific competition can drive community structure and function, and affect the distribution, demographic patterns, and behavioral strategies of co-occurring species (Chase et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2002\u003c/span\u003e, Chesson and Kuang \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2008\u003c/span\u003e, Vanak et al., \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Because large carnivore populations are primarily shaped by the abundance and availability of their main food items (Karanth et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Hayward \u0026amp; Kerley, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), prey represents a critical resource that coexisting carnivores can partition to reduce competition (Schoener, \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Hayward \u0026amp; Kerley, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Intra-guild competition intensity can vary spatio-temporally due to changes in prey availability, which can fluctuate due to a multitude of biotic and abiotic factors such as seasonality, droughts, climate change, or disease outbreaks (Walther et al., \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Duncan et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Zidon et al., \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). While numerous studies have investigated carnivore competition and resource partitioning, data is lacking in certain areas of sub-Saharan Africa, where largely intact carnivore guilds exist. Understanding how sympatric carnivores partition dietary resources in these systems is thus essential to understand carnivore ecology (Radloff and du Toit, \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) and inform conservation practices through vegetation and prey species management.\u003c/p\u003e\u003cp\u003eSome of the most diverse and multifaceted terrestrial assemblages of large carnivores occur in sub-Saharan Africa, with several species of large carnivores occurring sympatrically including African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) (hereafter lions), spotted hyenas (\u003cem\u003eCrocuta Crocuta\u003c/em\u003e), cheetahs (\u003cem\u003eAcinonyx jubatus\u003c/em\u003e), leopards (\u003cem\u003ePanthera pardus\u003c/em\u003e), and brown hyenas (\u003cem\u003eParahyaena brunnea\u003c/em\u003e). Lions and spotted hyenas are two of the most numerous large carnivores in this region where they overlap extensively in distribution (Hatton et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) and diel activity (Mills \u0026amp; Biggs, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Hayward \u0026amp; Hayward, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Patterson et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). To facilitate coexistence, lions and spotted hyenas have evolved disparate behavioral, social, and foraging strategies (P\u0026eacute;riquet et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Swanson et al., \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Lions are ambush predators, who hide under vegetation and pursue medium-large ungulate prey (Hayward and Kerley, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; P\u0026eacute;riquet et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), yet also hunt in open areas (Stander, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e1992\u003c/span\u003e) and scavenge opportunistically (Mole\u0026oacute;n et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Amor\u0026oacute;s et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Conversely, spotted hyenas are cursorial predators, typically chasing prey and using endurance for hunting success (Kruuk, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e1972\u003c/span\u003e; Mills, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1990\u003c/span\u003e) but also readily scavenge carcasses when available (Watts \u0026amp; Holekamp, \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Mole\u0026oacute;n et al., \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Additionally, spotted hyenas do successfully hunt calves or juvenile large ungulate species (Kruuk, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e1972\u003c/span\u003e; Holekamp et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Salnicki et al., \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eLions and spotted hyenas engage in kleptoparasitism; although spotted hyenas lose more of their kills to lions than lions lose to spotted hyenas (P\u0026eacute;riquet et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), though this kind of intra-guild competition is highly variable based on pride and clan size and structure (Trinkel and Kastberger, \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). With overlapping diel activity patterns, kleptoparasitism, and opportunistic scavenging, dietary overlap is expected to be high between lions and spotted hyenas with substantial variation across their shared geographic range. Previous studies report broad dietary niches for spotted hyenas (Vissia et al., \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), though they have been found to have similar (Hayward, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) or narrower (Hayward \u0026amp; Kerley, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) niche breadths compared to lions. However, spotted hyenas may reduce their niche overlap by preying on different prey size via different sex or age classes (P\u0026eacute;riquet et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Additionally, top prey items vary across studies for both species (e.g., Hayward, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e), and the role of seasonality in shaping dietary niche breadth and overlap remains poorly understood (Vissia et al., \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eFew studies have employed high-throughput sequencing methods like DNA metabarcoding to address these questions. DNA metabarcoding has been shown to have higher sensitivity and greater taxonomic resolution relative to conventional dietary studies that rely on morphological identification of undigested remains in scats (Stein et al., \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Galan et al., 2017). Morphological identification can introduce additional biases into dietary results through misidentification of closely related prey items, lack of detection for prey items with highly digestible tissues, and underestimating prey items (Monterosso et al., 2019). The conflicting findings in previous dietary studies, along with the limited application of molecular techniques, underscore the importance of assessing carnivore diet partitioning, especially between lions and spotted hyenas, to quantify dietary overlap in a rich carnivore community. Moreover, while little is known about the effects of seasonality on dietary niche overlap among large carnivores, understanding these dynamics is critical for advancing our knowledge of intra-guild resource partitioning and coexistence in shared landscapes.\u003c/p\u003e\u003cp\u003eOur goal in this study was to use DNA metabarcoding to quantify dietary patterns of sympatric lions and spotted hyenas in a dryland ecosystem in southern Africa. Specifically, our objectives were to 1) quantify the vertebrate diet composition of sympatric lions and spotted hyenas; 2) quantify seasonal variation in diet composition within and between each species; 3) assess dietary niche breadth for each species across seasons; and 4) evaluate interspecific dietary overlap to determine if there is evidence of potential niche partitioning and competition. We also formulated a series of predictions. First, we predicted that both species would have higher diversity in their diet during the wet season due to increased availability of young herbivores during the birthing season for many ungulate species, and animals dispersing across the landscape and away from predictable waterhole locations, forcing carnivores to be less selective for prey species. Second, based on scavenging activity and previous studies (Hayward, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Vissia et al., \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2023\u003c/span\u003e), we predicted that spotted hyenas would have higher dietary niche breadths than lions during both seasons. Lastly, we predicted that both species would have high dietary overlap during both seasons because of kleptoparasitism and opportunistic scavenging.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStudy Area\u003c/h2\u003e\u003cp\u003eThis study was conducted in the Greater Etosha Landscape (GEL) encompassing Etosha National Park (\u0026ldquo;Etosha\u0026rdquo;), the Ongava Game Reserve (\u0026ldquo;Ongava\u0026rdquo;), and King Nehale communal conservancy (\u0026ldquo;King Nehale\u0026rdquo;) in north-central Namibia (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) (Turner et al., \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Etosha is a 22,900 km\u003csup\u003e2\u003c/sup\u003e fenced protected area, although the fence is permeable to wildlife (Naha et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Melton, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Ongava is a fenced 300 km\u003csup\u003e2\u003c/sup\u003e private game reserve on the southern border of Etosha. King Nehale is a 508 km\u003csup\u003e2\u003c/sup\u003e communal conservancy located in the Oshikoto region, bordering the northeast corner of Etosha. One main source of income for people living in the conservancy is keeping livestock, including cattle, goats, and sheep (Nakanyala et al., \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The GEL is located within the semi-arid weather zone with an average annual rainfall of 380 mm (De Beer et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Atlas of Namibia, 2022). Despite fencing around Etosha, the prey and predator guild composition in all these areas is very similar and together they form part of the same larger scale ecosystem, including movement of carnivores between Ongava and Etosha, and more so between King Nehale and Etosha (Melton, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). The main vegetation types in Etosha are grasslands, steppe, shrubland, Mopane (\u003cem\u003eColophospermum mopane\u003c/em\u003e) tree savanna, and mixed tree savanna (Le Roux et al., \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1988\u003c/span\u003e). The landcover on Ongava is classified as Karstveld, with Mopane shrubland and woodland being the dominant vegetation type (~\u0026thinsp;70%) and other areas with savannah-like vegetation (~\u0026thinsp;30%, Berry and Loutit, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). During the wet season (November-April), rainfall provides additional temporal water sources, aside from the permanent artificial water sources across the landscape (Engert, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Berezin et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Both Etosha and Ongava are tourist destinations for wildlife-based viewing and support a broad diversity of carnivore and ungulate species [e.g., black-faced impala (\u003cem\u003eAepyceros melampus petersi\u003c/em\u003e), springbok (\u003cem\u003eAntidorcas marsupialis\u003c/em\u003e), blue wildebeest (\u003cem\u003eConnochaetus taurinus\u003c/em\u003e), plains zebra (\u003cem\u003eEquus quagga burchelli\u003c/em\u003e), gemsbok (\u003cem\u003eOryx gazella\u003c/em\u003e)], including megaherbivores [e.g, African elephant (\u003cem\u003eLoxodonta africana\u003c/em\u003e), southern Angolan giraffe (\u003cem\u003eGiraffa giraffa angolensis\u003c/em\u003e), southern black rhinoceros (\u003cem\u003eDiceros bicornis bicornis\u003c/em\u003e)]. However, prey species\u0026rsquo; composition and abundance differ between Ongava and Etosha, and across the different regions of Etosha (west, central, and east) (Trinkel, \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Kilian and Kolberg, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eSample Collection\u003c/h3\u003e\n\u003cp\u003eWe collected 164 dried or fresh fecal samples [spotted hyena (n\u0026thinsp;=\u0026thinsp;32) and lion (n\u0026thinsp;=\u0026thinsp;132)] from October 2021 through March 2024 around Etosha, Ongava, and King Nehale. When divided by location, we collected 72 samples in Etosha [spotted hyena (n\u0026thinsp;=\u0026thinsp;28) and lion (n\u0026thinsp;=\u0026thinsp;44)], three samples in King Nehale [spotted hyena (n\u0026thinsp;=\u0026thinsp;2) and lion (n\u0026thinsp;=\u0026thinsp;1)], and 88 samples in Ongava [spotted hyena (n\u0026thinsp;=\u0026thinsp;3) and lion (n\u0026thinsp;=\u0026thinsp;82)]. Most samples (n\u0026thinsp;=\u0026thinsp;140) were collected opportunistically from the ground while driving along roads, around waterholes, and near carnivore kill sites within our study area (approved by Namibian Ministry of Environment, Forestry, and Tourism under permit #AN202101004). For these samples, whole fecal samples were collected into a paper bag and dried naturally before being subsampled for DNA extraction, which included both the external surface and internal matrix to capture both consumer and prey DNA. Additionally, 24 samples [lions (n\u0026thinsp;=\u0026thinsp;19), spotted hyenas (n\u0026thinsp;=\u0026thinsp;5)] were collected during carnivore immobilizations as part of ongoing research (Appendix S1; Text S1). These samples were manually retrieved from the immobilized animal, placed into an open plastic vial and stored in a paper bag for natural drying. Due to the small amount collected directly from immobilized animals, these samples were not subsampled. All immobilizations were performed by veterinarians registered with the Namibian Veterinary Council and the Ministry of Environment, Forestry, and Tourism and were approved by the Namibian Ministry of Environment, Forestry, and Tourism (permit #AN202101004) as well as the University of Georgia Institutional Animal Care And Use Committee under protocols A2024 05-009-06 and A2021 04-013-Y3-A11.\u003c/p\u003e\n\u003ch3\u003eLaboratory Analyses and Data Curation\u003c/h3\u003e\n\u003cp\u003eAll samples were shipped from Namibia to the United States in July 2024 and DNA was extracted from feces using DNeasy PowerSoil Pro DNA Kits (QIAGEN) in August 2025. Jonah Ventures, LLC conducted DNA amplification, sequencing, and bioinformatics for DNA metabarcoding (Appendix 1; Text S2). This technique analyzes DNA within scats using high-throughput sequencing using small, highly variable universal primers to identify prey species and confirm predator species (Pompanon et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Xiong et al., 2017). To characterize prey DNA from carnivore fecal samples, we amplified a portion of the 12S rRNA gene using the primer pair 12SVertF (forward 5\u0026prime;- ACTGGGATTAGATACCCYACTATG \u0026minus;\u0026thinsp;3\u0026prime;) and 12SVertR (reverse 5\u0026prime;- GAGRRTGACGGGCGGTDT \u0026minus;\u0026thinsp;3\u0026prime;) (Evans et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Both forward and reverse primers contained a 5\u0026rsquo; adaptor sequence to allow for subsequent indexing and Illumina sequencing. Exact sequence variants (ESVs) were identified using the UNOISE3 algorithm (Edgar, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2016a\u003c/span\u003e) implemented in VSEARCH (Rogens et al., 2016), retaining only those with \u0026ge;\u0026thinsp;4 read counts and \u0026le;\u0026thinsp;1 expected error. We assigned taxa of ESVs using a custom reference database consisting of publicly available sequences on NCBI GenBank (Benson et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) and Jonah Ventures voucher sequences records, accepting the reference taxonomy for any taxonomic level with \u0026gt;\u0026thinsp;90% agreement within 1% of the top hits. We discarded any reads of taxa not occurring in the study region, and to limit the effects of contamination, only prey species that consisted of more than 1% of total prey sequences were retained (Shively et al., \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Because host DNA typically dominates the sequencing dataset (Forin-Wiart et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), we were able to confirm predator species for each sample.\u003c/p\u003e\n\u003ch3\u003eData Analyses\u003c/h3\u003e\n\u003cp\u003eWe conducted all analyses and produced figures using R version 4.1.1 (R Core Team, \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). We summarized the data with both frequency of occurrence (FOO) and relative read abundance (RRA) at the species level, and calculated prey species richness for each carnivore species by season (wet and dry). To calculate FOO, we divided the number of occurrences of a prey species by the total number of carnivore scat samples and expressed FOO as a percentage. We calculated RRA for each prey species as the read count (the number of DNA sequence reads) for that the given prey species divided by the total number of reads in the carnivore scat sample, also expressed as a percentage.\u003c/p\u003e\u003cp\u003eTo examine diet composition (beta-diversity) of vertebrate prey species, we conducted separate three-dimensional non-metric multidimensional scaling (NMDS) analyses with 1000 permutations to visualize trends in variation between seasons and carnivore species. We used two complementary approaches to evaluate dietary similarity between and within carnivore species. First, we applied a non-parametric Analysis of Similarities (ANOSIM) with binary Jaccard dissimilarities to test whether prey composition within carnivore scats differed between lions and hyenas within each season. Because ANOSIM evaluates overall compositional differences but does not quantify the degree of overlap, we also calculated Pianka\u0026rsquo;s index of niche overlap (Pianka, \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e1973\u003c/span\u003e) based on FOO. To account for unequal sample sizes, we generated 95% bootstrap confidence intervals by stratified resampling of carnivore scats for both species (2,000 iterations). Together, these analyses allowed us to assess both whether diets differed significantly between carnivore species and the extent of dietary niche overlap, providing a more complete picture of dietary similarity across seasons. We used the vegan package in R (Oksanen et al., \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) to conduct the NMDS and ANOSIM and spaa package in R (Zhang, \u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) to determine Pianka\u0026rsquo;s index.\u003c/p\u003e\u003cp\u003eWe calculated dietary niche breadth (\u003cem\u003eBa\u003c/em\u003e) for each carnivore species in both seasons by using Hurlbert\u0026rsquo;s standardized version of Levin's measure (Hurlbert, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e1978\u003c/span\u003e; Smith, \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e1982\u003c/span\u003e) for FOO proportions of prey. To account for differences in sample size across carnivore species and seasons, we applied a bootstrap approach with 2,000 iterations. For each iteration, all samples were resampled with replacement within each carnivore species\u0026ndash;season group, and standardized niche breadth was recalculated. Broader dietary niche breadth is indicated by a higher \u003cem\u003eBa\u003c/em\u003e while a narrow dietary niche breadth is indicated by a low \u003cem\u003eBa\u003c/em\u003e (ranging from 0\u0026ndash;1). We did this overall with all data pooled, and separately for each study site [Etosha (including King Nehale) versus Ongava].\u003c/p\u003e\u003cp\u003eTo evaluate completeness of our sampling efforts and prey richness, we generated sample-based rarefaction and extrapolation curves for each carnivore species and season using the specaccum function in the vegan package (Oksanen et al., 2022). Scat samples were converted to presence\u0026ndash;absence matrices of prey taxa, and 1,000 random permutations were used to estimate mean prey richness and associated 95% confidence intervals. We further estimated asymptotic richness with the Chao1 estimator (specpool function) and plotted these estimates (\u0026plusmn;\u0026thinsp;standard error) alongside rarefaction curves to assess whether observed prey detections approached estimated richness. For all analyses we pooled data from all study sites to generate broad inferences about carnivore diet, and we also visually examined diet composition between Etosha and King Nehale versus Ongava.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eDiet overview\u003c/h2\u003e\u003cp\u003eFrom the 164 samples that were sequenced, 98 had detectable vertebrate prey items and were included in analysis [spotted hyena (n\u0026thinsp;=\u0026thinsp;29; dry season\u0026thinsp;=\u0026thinsp;17, wet season\u0026thinsp;=\u0026thinsp;12) and lion (n\u0026thinsp;=\u0026thinsp;69; dry season\u0026thinsp;=\u0026thinsp;54, wet season\u0026thinsp;=\u0026thinsp;15)]. Overall, we detected 9 families and 17 genera encompassing 19 vertebrate prey species (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). For spotted hyenas, the total read count was 127640 with a mean read count of 3868 per sample (\u0026plusmn;\u0026thinsp;4461 SD). For lions, the total read count was 107965 with a mean read count of 1367 per sample (\u0026plusmn;\u0026thinsp;2733 SD).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePercent frequency of occurrence (%FOO; number of fecal samples containing each prey species divided by total number of fecal samples and expressed as a percentage) and Relative Read Abundance (RRA%; total number of reads of each vertebrate prey species divided by total number of vertebrate reads and expressed as a percentage) of vertebrate species (12S rRNA) detected in the diet of African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) based on season (dry\u0026thinsp;=\u0026thinsp;May-October; wet\u0026thinsp;=\u0026thinsp;November-April) in the Greater Etosha Landscape; \u0026ldquo;n\u0026rdquo; represents number of samples containing that prey species for that carnivore species\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCarnivore - Season\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePrey Species\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003en\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAverage RRA (%)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eFOO (%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"14\" rowspan=\"15\"\u003e\u003cp\u003eLion \u0026ndash;\u003c/p\u003e\u003cp\u003eDry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBaboon\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.69\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlack rhinoceros\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.39\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlack-faced impala\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.99\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.69\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlue wildebeest\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.56\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCommon eland\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.86\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e13.56\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGemsbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e25.42\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAngolan giraffe\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.08\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHare spp.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.29\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.39\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGreater kudu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.08\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eMountain zebra\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.39\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePlains zebra\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e6.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10.17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRed hartebeest\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3.95\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.39\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSpringbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e5.93\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.78\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTurner's thick-toed gecko\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.69\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eWhite rhinoceros\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.69\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"9\" rowspan=\"10\"\u003e\u003cp\u003eLion \u0026ndash;\u003c/p\u003e\u003cp\u003eWet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlack-faced impala\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e14.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlue wildebeest\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCommon eland\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGemsbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e13.90\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAngolan giraffe\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e14.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHare spp.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e7.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGreater kudu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e9.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePlains zebra\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e9.76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePython spp.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e4.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRed hartebeest\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e9.69\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"7\" rowspan=\"8\"\u003e\u003cp\u003eSpotted Hyena - Dry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlack rhinoceros\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlack-faced impala\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e11.76\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCow\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCommon eland\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGemsbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e17.65\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGreater kudu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePlains zebra\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e29.41\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSpringbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e17.65\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\" morerows=\"8\" rowspan=\"9\"\u003e\u003cp\u003eSpotted Hyena - Wet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eBlack rhinoceros\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e3.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCow\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.25\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCommon eland\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.25\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGemsbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e12.1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e18.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eAngolan giraffe\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e13.07\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGoat\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15.7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e12.5\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eGreater kudu\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e10.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.25\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePlains zebra\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e14.13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e18.75\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSpringbok\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e15.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e6.25\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eSpotted hyena diet\u003c/h3\u003e\n\u003cp\u003eFrom 29 sequenced spotted hyena samples, plains zebra was the most frequently consumed prey species in the dry season (FOO\u0026thinsp;=\u0026thinsp;29.4%) followed by gemsbok (\u003cem\u003eOryx gazella\u003c/em\u003e) and springbok (FOO\u0026thinsp;=\u0026thinsp;17.7% each). In the wet season, spotted hyenas equally consumed plains zebras and gemsbok (FOO\u0026thinsp;=\u0026thinsp;18.8% each, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Overall, prey species richness was 10 and included two domestic livestock species, cow (\u003cem\u003eBos taurus\u003c/em\u003e; n\u0026thinsp;=\u0026thinsp;2) and goat (\u003cem\u003eCapra hircus\u003c/em\u003e; n\u0026thinsp;=\u0026thinsp;2). Both domestic prey species were found in samples near or outside of northeast Etosha (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). ANOSIM revealed no seasonal differences in diet composition for spotted hyenas (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eANOSIM results comparing prey composition between African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) in the Greater Etosha Landscape. Comparisons include between seasons (wet\u0026thinsp;=\u0026thinsp;November \u0026ndash; April; dry\u0026thinsp;=\u0026thinsp;May \u0026ndash; October) within each species, seasons between species, and overall between species across all samples. R-statistic and p-values are provided for each comparison\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e\u003cp\u003eSeasonality within species (wet vs.dry)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eR-stat\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003ep-value\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSpotted Hyena\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-0.024\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.73\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-0.008\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.58\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e\u003cp\u003eSeasonality between species\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.021\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-0.018\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.67\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e\u003cp\u003eOverall between species\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLion vs. Spotted Hyena\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.014\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.17\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eWhen we separated out by study site (Etosha and King Nehale versus Ongava), during the dry season, plains zebra and springbok were the most consumed prey items in Etosha (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e) while gemsbok and kudu (\u003cem\u003eTragelaphus strepsiceros\u003c/em\u003e) were the only consumed prey items in Ongava (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). During the wet season, spotted hyenas primarily consumed plains zebra and gemsbok in Etosha (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e).\u003c/p\u003e\n\u003ch3\u003eLion diet\u003c/h3\u003e\n\u003cp\u003eFrom 69 sequenced samples, lions consumed a wide variety of species but primarily large ungulates such as gemsbok (FOO\u0026thinsp;=\u0026thinsp;25.4%), followed by blue wildebeest and eland (\u003cem\u003eTaurotragus oryx\u003c/em\u003e) (FOO\u0026thinsp;=\u0026thinsp;13.6% each), then plains zebra (FOO\u0026thinsp;=\u0026thinsp;10.2%) during the dry season. In the wet season lions consumed eland, gemsbok, and plains zebra equally (FOO\u0026thinsp;=\u0026thinsp;15% each), followed by black-faced impala, blue wildebeest, giraffe, and hare species (FOO\u0026thinsp;=\u0026thinsp;10% each, Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Overall, prey richness was 16 and ANOSIM revealed no significant seasonal differences in diet composition for lions (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eSeparated out by study site, during the dry season, lions consumed springbok most frequently in Etosha, followed by giraffe, plains zebra, eland, red hartebeest, and black rhinoceros equally (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). During the dry season on Ongava, lions consumed gemsbok most frequently, followed by blue wildebeest, eland, and plains zebra (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e). During the wet season in Etosha, lions primarily consumed giraffe and plains zebra equally followed by eland, red hartebeest, blue wildebeest, kudu, and python species equally (Fig. \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). During the wet season on Ongava, lions, consumed gemsbok, eland, black-faced impala, and hare spp. equally, followed by blue wildebeest and plains zebra (Fig. \u003cspan refid=\"MOESM2\" class=\"InternalRef\"\u003eS2\u003c/span\u003e).\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eDietary niche breadth\u003c/h2\u003e\u003cp\u003eOverall, during the wet and dry seasons, lions exhibited broader dietary niche breadth than spotted hyenas [lions (\u003cem\u003eBa\u003c/em\u003e: dry\u0026thinsp;=\u0026thinsp;0.40, wet\u0026thinsp;=\u0026thinsp;0.45); hyenas (\u003cem\u003eBa\u003c/em\u003e: dry\u0026thinsp;=\u0026thinsp;0.28, wet\u0026thinsp;=\u0026thinsp;0.38)]. Both species had broader dietary niche breadths during the wet season compared to the dry season (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). In Etosha, lions exhibited broader dietary niche breadth than spotted hyenas in the dry season (\u003cem\u003eBa\u003c/em\u003e: lions\u0026thinsp;=\u0026thinsp;0.5, hyenas\u0026thinsp;=\u0026thinsp;0.29), but spotted hyenas had slightly broader dietary niche breadths in the wet season (\u003cem\u003eBa\u003c/em\u003e: lions\u0026thinsp;=\u0026thinsp;0.47, hyenas\u0026thinsp;=\u0026thinsp;0.44). Lions had broader dietary niche breadths in the dry season, while spotted hyenas were broader in the wet season (Fig. S3). In Ongava, lions had similar dietary niche breadths across seasons (\u003cem\u003eBa\u003c/em\u003e: dry\u0026thinsp;=\u0026thinsp;0.417, wet\u0026thinsp;=\u0026thinsp;0.414), and broader dietary niche breadth compared to spotted hyenas in the dry season (\u003cem\u003eBa\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.09) (Fig. S4).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\u003ch2\u003eInterspecific dietary variation\u003c/h2\u003e\u003cp\u003eWe found no differences in dietary composition for either carnivore species overall (all data pooled) or across season. The NMDS ordination for dietary overlap (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e) had a stress value of 0 in the wet season and \u0026lt;\u0026thinsp;0.001 in the dry season. While a stress value of or near 0 is uncommon, it can occur in small or low-dimensional datasets but is under the threshold value (stress\u0026thinsp;\u0026lt;\u0026thinsp;0.2) as suggested by Clarke (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1993\u003c/span\u003e) for an interpretable ordination. Pairwise ANOSIM comparisons revealed no significant differences (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) in diet composition between lions and spotted hyenas within either season or overall. R statistics were close to zero or negative, indicating strong dietary overlap among these carnivores across seasons and overall (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Pianka\u0026rsquo;s index revealed moderate dietary overlap between lions and spotted hyenas overall (O\u0026thinsp;=\u0026thinsp;0.75). Seasonal analyses showed that overlap between species was slightly higher in the dry season (O\u0026thinsp;=\u0026thinsp;0.68) than in the wet season (O\u0026thinsp;=\u0026thinsp;0.66). Within species, lions exhibited greater dietary similarity across wet and dry seasons (O\u0026thinsp;=\u0026thinsp;0.84) compared to spotted hyenas (O\u0026thinsp;=\u0026thinsp;0.77) (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003ePianka\u0026rsquo;s index of dietary overlap (O) between African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e). Overlap values are shown with 95% confidence intervals (CIs) based on bootstrap resampling (2,000 iterations). Results are presented for seasonal comparisons within species (wet vs. dry), between species within each season, and overall between species across all samples\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e\u003cp\u003eSeasonality within species (wet vs.dry)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e\u003cb\u003eO\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e\u003cb\u003e95% CI\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSpotted Hyena\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.34\u0026ndash;0.85\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.84\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.53\u0026ndash;0.88\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e\u003cp\u003eSeasonality between species\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDry\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.68\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.36\u0026ndash;0.80\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWet\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.31\u0026ndash;0.74\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c3\" namest=\"c1\"\u003e\u003cp\u003eOverall between species\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLion vs. Spotted Hyena\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.54\u0026ndash;0.83\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003eRarefaction curves\u003c/h2\u003e\u003cp\u003eRarefaction curves revealed that prey species richness in lion scats approached asymptotic levels, particularly in the dry season, where observed richness (15 taxa) closely matched Chao1 estimates (17.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6). In the wet season, lion richness was lower (10 taxa, 11.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7), suggesting some rare prey may not have been captured. Spotted hyena curves indicated lower observed richness overall (dry\u0026thinsp;=\u0026thinsp;8, wet\u0026thinsp;=\u0026thinsp;9 taxa), with estimates suggesting additional undetected prey species, particularly in the dry season (Chao1\u0026thinsp;=\u0026thinsp;15.5\u0026thinsp;\u0026plusmn;\u0026thinsp;11.0), though uncertainty was high due to small sample size (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eDetermining how large carnivores partition dietary resources is critical for understanding intra-guild interactions and coexistence. Using DNA metabarcoding, we characterized the diets of lions and spotted hyenas in the GEL and found high dietary overlap between both species, regardless of the season. Despite differences in prey species richness and niche breadth between carnivore diets, there were no significant seasonal or interspecific differences in prey species composition. These findings suggest that in this landscape, resource partitioning among large carnivores may occur by targeting different sex or age classes of prey items or through mechanisms other than diet. Collectively, these data contribute to our overall understanding of dietary patterns of sympatric lions and spotted hyenas in a semi-arid ecosystem.\u003c/p\u003e\u003cp\u003eSpecies with broad dietary niche breadths are typically considered dietary generalists while those with narrow dietary niche breadths are considered specialists (Carvalho \u0026amp; Cardoso, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). In our study, lions had a wider dietary niche breadth during both seasons compared to spotted hyenas; however, our findings demonstrate that lions and spotted hyenas exhibit broadly similar diets with consistently high overlap, suggesting that these two apex predators rely heavily on a shared prey base. When comparing dietary niche breadth between species, lions and spotted hyenas are reported to have similar dietary niche breadths, which aligns with our results (Hayward, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Hayward and Kerley, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2008\u003c/span\u003e, reviewed in P\u0026eacute;riquet et al., \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The comparison between seasons supported our prediction that both lions and spotted hyenas had broader dietary niche breadths in the wet season compared to the dry. This difference in seasonality has been previously documented in African mammal diets (Vissia et al., \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) perhaps because some prey species are more abundant and more easily accessible (Trinkel, \u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Pereira et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) during the birthing season or because rainfall leads to prey dispersing across the landscape. As a result, predators can no longer rely on high prey densities near predictable waterhole locations and may be less selective, consuming whatever prey they encounter.\u003c/p\u003e\u003cp\u003eThe results confirm our prediction that there would be high dietary overlap between both carnivore species, likely enhanced by kleptoparasitism and scavenging behavior. The overall Pianka\u0026rsquo;s index value (0.75) indicates substantial dietary similarity, and this pattern was maintained across seasons, with only minor fluctuations between the wet and dry periods. Apart from domestic livestock species, all prey species detected in the spotted hyena diet were also found in the lion diet. Extensive dietary overlap may indicate occurrences of kleptoparasitism by lions and spotted hyenas, which previously have been reported (Trinkel and Kastberger, \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Additionally, lions and spotted hyenas are documented competitors that consume similar prey items (Hayward, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Hayward and Kerley, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) and exhibit comparable temporal activity patterns (Mills \u0026amp; Biggs, \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Hayward \u0026amp; Hayward, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Patterson et al., \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Thus, our findings indicate that lions and spotted hyenas in Etosha may partition their prey resources by age or sex class, or spatially to co-exist (Patterson et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) and we recommend further studies to quantify prey partitioning across demographic classes and spatial scales to better understand mechanisms of niche differentiation and coexistence among large carnivores.\u003c/p\u003e\u003cp\u003eIn a review of spotted hyena diet across several countries, Hayward (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) determined that gemsbok was the most frequently consumed prey species where it occurred, and plains zebra and springbok were typically consumed infrequently. Conversely, other studies (Gasaway et al., \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Trinkel, \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) found plains zebra and springbok were the main prey species for spotted hyenas in some areas of Etosha, while Berry (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1981\u003c/span\u003e) determined wildebeest were the main prey species in Etosha. Fester et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) found gemsbok and springbok were the main prey items in the Namib desert of southwest Namibia. In our study, spotted hyenas most frequently consumed plains zebras, gemsbok, and springbok, respectively. As such, spotted hyenas may target gemsbok but also consume plains zebras and springbok in the GEL due to their high abundance and availability compared to other ungulates. For example, in 2015, the Etosha density of gemsbok was estimated at 0.22 individuals/km\u003csup\u003e2\u003c/sup\u003e while plains zebra was 0.77 individuals/km\u003csup\u003e2\u003c/sup\u003e and springbok were 0.56 individuals/km\u003csup\u003e2\u003c/sup\u003e (Kilian and Kolberg, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). As plains zebra and springbok are grazers (Kingdon, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), and gemsbok are mixed feeders that primarily graze (Sponheimer et al., \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Lehmann et al., \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), these results also align with Patterson et al. (\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) who found that spotted hyenas in Etosha select for areas of high grass cover during both seasons. Collectively, these studies indicate that in the GEL, spotted hyenas are likely hunting in open, grassy areas and primarily consuming grazing prey species.\u003c/p\u003e\u003cp\u003eThe range of prey items for lions showed no strong selection for either browsers or grazers, as both (browsers: giraffe; mixed: eland, gemsbok; grazers: plains zebra, blue wildebeest, Kingdon, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) were found as lion top prey species. Our results align with findings from Hayward and Kerley (\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) who found lions\u0026rsquo; primary prey species were gemsbok, blue wildebeest, giraffe, eland, and zebra across several countries and Berry (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1981\u003c/span\u003e) who found lions\u0026rsquo; primary prey items in Etosha were gemsbok, wildebeest, and plains zebra. Consistent with other findings, lions opt for large prey species but will also consume small prey opportunistically (Hayward and Kerley, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Davidson et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Barnardo et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This supports the optimal foraging theory, which suggests that a predator can distinguish among prey of differing profitability and typically select the most profitable types (Krebs, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e1978\u003c/span\u003e). Large-bodied grazers were consumed more during the dry season in our study, which may indicate that lions were opting for higher caloric returns from large prey species to sustain themselves during higher periods of stress (Hayward and Kerley, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). Additionally, in the dry season herbivores congregate at fixed waterhole locations in the GEL, increasing predator access to a higher density of potential prey. Conversely, in the wet season lions appear to consume fewer species, with several at higher frequencies relative to the dry season (e.g., hare spp., black-faced impala, giraffe), potentially reflecting opportunistic foraging or scavenging due to greater prey dispersion.\u003c/p\u003e\u003cp\u003eWe found three spotted hyena samples with domestic livestock species (cow and goat). All samples were collected near or outside of Etosha in the northeast area (King Nehale communal conservancy) where livestock are abundant and spotted hyenas often leave the park and enter onto communal land (Naha et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Melton, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Patterson et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), sometimes resulting in retaliatory killings of carnivores after livestock depredation events (Stander, \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Goelst, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2018\u003c/span\u003e, Naha et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Unfortunately, there is little information on the extent and demography of spotted hyenas killed in anthropogenic areas surrounding Etosha, but local farmers report frequent sightings of spotted hyenas on their farms (Lendelvo et al., \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). These findings contribute to our overall understanding of livestock depredation in a high human-carnivore conflict area.\u003c/p\u003e\u003cp\u003eThe rarefaction analyses highlight important differences in sampling completeness between lions and spotted hyenas, which help contextualize niche overlap between lions and spotted hyenas, as well as niche breadth estimates. We could not detect prey items in 66 samples due to predator DNA swamping, which occurs when host DNA is present in much higher concentrations than prey DNA (Shi et al., \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). For lions, both wet- and dry-season rarefaction curves approached asymptotic levels, and Chao1 estimates suggested that only a small number of prey species were likely undetected, indicating that our dietary characterization for lions is robust and provides a reliable picture of seasonal prey use. In contrast, the rarefaction curve for spotted hyenas showed lower observed richness and substantially higher Chao1 estimates, particularly in the dry season where uncertainty in prey species detection was large, reflecting potential under-sampling. This limitation suggests that while lions and spotted hyenas appear to have high dietary overlap, the true extent of spotted hyena prey use may be broader than captured in this study. Consequently, the high dietary overlap we observed may partially reflect more complete sampling of lion diets compared to spotted hyenas. Nonetheless, the combination of dietary niche overlap and breadth results indicates that both species rely heavily on shared prey resources, with lions exhibiting more stable and well-characterized prey use across seasons. Future work that includes expanded spotted hyena sampling could help clarify whether spotted hyena dietary breadth converges more strongly with or diverges from that of lions under different seasonal conditions.\u003c/p\u003e\u003cp\u003eWe found notable differences in frequently consumed prey species and dietary niche breadth for both lions and spotted hyenas between study sites, likely reflecting variation in prey abundance and community composition within the sampled areas. Although both Etosha and Ongava are protected areas, they are managed by different entities and are different sizes, which may influence wildlife distributions and prey availability. However, the small number of spotted hyena samples from Ongava (n\u0026thinsp;=\u0026thinsp;2) limited our ability to accurately characterize their dietary niche breadth at that site. Additionally, because sampling did not encompass the entire extent of Etosha, we were unable to provide a comprehensive synthesis of lion and spotted hyena diets across the entire GEL. Despite these limitations, this study provides valuable insights into the diets of lions and spotted hyenas where dietary data, particularly derived from high-resolution DNA metabarcoding, are lacking.\u003c/p\u003e\u003cp\u003eOur study contributes to the area of emerging research using DNA metabarcoding analysis to compare diets of large, sympatric carnivores. The results revealed strong dietary overlap and limited niche partitioning among lions and spotted hyenas across both wet and dry seasons. Our findings suggest that prey availability and opportunistic feeding behavior may be the primary drivers of diet composition in this well-preserved carnivore guild. Despite differences in dietary niche breadth, both species showed high overlap in key prey taxa, perhaps due to opportunistic scavenging and kleptoparasitism. These findings emphasize the importance of maintaining abundant ungulate populations to support diverse predator communities. Importantly, the detection of domestic livestock in spotted hyena diets highlights the potential for human\u0026ndash;wildlife conflict, emphasizing the need for continued management interventions around Etosha. Further diet monitoring is essential for detecting shifts in predator\u0026ndash;prey dynamics in response to environmental change, prey population fluctuations, or increased anthropogenic pressures. Future research should also explore the spatial and temporal aspects of scavenging and interspecific interactions to better inform adaptive management and conservation strategies in protected areas and surrounding multi-use landscapes.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eCompeting interests:\u003c/h2\u003e\u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding:\u003c/h2\u003e\u003cp\u003eThis work was supported by University of Georgia and the US Department of Energy Office of Environmental Management Award Number DE-EM0005228 to the University of Georgia Research Foundation. Disclaimer: This manuscript was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information disclosed, or represents that its use not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not constitute or imply its endorsement, recommendation, or favoring by the United States Government or any agency thereof. The views and opinions of the authors expressed herein do not necessarily state or reflect those of the United States Government or any agency thereof.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eJessica Patterson, James Beasley, and Stephanie Periquet-Pearce contributed to the study conception and design. Material preparation and data collection were performed by all authors. Date analyses and the first draft of the manuscript were completed by Jessica Patterson. All authors commented on previous versions of the manuscript and approved the final manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe thank the Ongava Game Reserve for providing some equipment and staff assistance. We thank the field technicians for their work in helping to collect and process scat/fecal samples. Thank you to the University of Georgia and the US Department of Energy Office of Environmental Management for funding assistance.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe datasets generated and analysed during the current study are available in the NCBI Sequence Read Archive repository at [https://www.ncbi.nlm.nih.gov/sra/PRJNA1357869](https:/www.ncbi.nlm.nih.gov/sra/PRJNA1357869) .\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAmor\u0026oacute;s, M., Gil‐S\u0026aacute;nchez, J.M., L\u0026oacute;pez‐Pastor, B.D.L.N. and Mole\u0026oacute;n, M. 2020. Hyaenas and lions: how the largest African carnivores interact at carcasses. \u003cem\u003eOikos\u003c/em\u003e, 129(12): 1820-1832.\u003c/li\u003e\n\u003cli\u003eAtlas of Namibia Team. 2022. 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Zebra migration strategies and anthrax in Etosha National Park, Namibia. \u003cem\u003eEcosphere\u003c/em\u003e, 8(8): e01925.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"African lions, diet composition, dietary niche breadth, interspecific competition, resource partitioning, spotted hyenas","lastPublishedDoi":"10.21203/rs.3.rs-7990153/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7990153/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eUnderstanding how large carnivores partition dietary resources is essential for assessing intra-guild competition and informing conservation strategies. In this study, we used DNA metabarcoding of scats to quantify and compare the diets of sympatric African lions (\u003cem\u003ePanthera leo\u003c/em\u003e) and spotted hyenas (\u003cem\u003eCrocuta crocuta\u003c/em\u003e) across wet and dry seasons in the Greater Etosha Landscape of Namibia. Across 98 scat samples, we identified 19 vertebrate prey species. Overall, large ungulates dominated both carnivores\u0026rsquo; diets. For lions, the most frequent prey items included gemsbok (\u003cem\u003eOryx gazella\u003c/em\u003e), common eland (\u003cem\u003eTaurotragus oryx\u003c/em\u003e), plains zebra (\u003cem\u003eEquus quagga burchelli\u003c/em\u003e), and blue wildebeest (\u003cem\u003eConnochaetus taurinus\u003c/em\u003e). For spotted hyenas, the most frequent prey items were plains zebra, gemsbok, springbok (\u003cem\u003eAntidorcas marsupialis\u003c/em\u003e), and black rhinoceros (\u003cem\u003eDiceros bicornis bicornis\u003c/em\u003e). Despite differing dietary niche breadth, with lions exhibiting the broadest across both seasons, diet composition was similar between species and seasons. These results confirm a high diet overlap and limited resource partitioning both within and among these large carnivore species across seasons, likely facilitated by opportunistic scavenging and kleptoparasitism. Both species exhibited broader dietary niche breadths during the wet season, likely reflecting increased prey availability and dispersion. Ongoing monitoring of carnivore diets using molecular tools, which provides a more accurate and comprehensive identification of diet items than manual sorting, will be essential for detecting changes in resource use and interspecific interactions in response to shifting environmental conditions and anthropogenic pressures.\u003c/p\u003e","manuscriptTitle":"Revealing Seasonal Dietary Niche Overlap Among Sympatric Large Carnivores using DNA Metabarcoding","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-26 16:04:40","doi":"10.21203/rs.3.rs-7990153/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-12-22T05:54:08+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-10T04:49:52+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-04T22:28:39+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"333915805512745477194129090686326205958","date":"2025-11-21T10:10:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"186499056832541225312642635568251851856","date":"2025-11-18T18:51:08+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"54819571020717858714702071131446504099","date":"2025-11-18T18:48:38+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-18T18:35:49+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-17T17:24:57+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-11-17T12:42:55+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-13T13:47:35+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-11-13T13:44:07+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5fbaf570-644a-484a-a4c7-180bf0fcfa2e","owner":[],"postedDate":"November 26th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":58519553,"name":"Biological sciences/Ecology"},{"id":58519554,"name":"Earth and environmental sciences/Ecology"},{"id":58519555,"name":"Biological sciences/Zoology"}],"tags":[],"updatedAt":"2026-03-16T16:06:22+00:00","versionOfRecord":{"articleIdentity":"rs-7990153","link":"https://doi.org/10.1038/s41598-026-43423-w","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-03-15 15:59:18","publishedOnDateReadable":"March 15th, 2026"},"versionCreatedAt":"2025-11-26 16:04:40","video":"","vorDoi":"10.1038/s41598-026-43423-w","vorDoiUrl":"https://doi.org/10.1038/s41598-026-43423-w","workflowStages":[]},"version":"v1","identity":"rs-7990153","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7990153","identity":"rs-7990153","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}